Cloning and expression of the pneumococcal neuraminidase gene in Escherichia coli

Anne M. Berry, James C. Paton, Eric M. Glare, David Hansman, David E.A. Catcheside

    Research output: Contribution to journalArticlepeer-review

    24 Citations (Scopus)


    A gene bank of Sau3AI-generated Streptococcus pneumoniae DNA fragments was constructed in Escherichia coli K-12 by cloning into the BamHI site of the cosmid vector pHC79. One clone capable of cleaving the fluorogenic neuraminidase substrate 2'-(4-methylumbelliferyl)-α-d-N-acetyl-neuraminic acid was isolated. This activity was inhibited by treatment with a mouse antiserum raised against purified pneumococcal neuraminidase. The recombinant plasmid purified from this clone (designated pJCP301) contained approximately 3.0 kb of pneumococcal DNA. Western-blot analysis indicated that E. coli K-12[pJCP301] produced a 98-kDa polypeptide which reacted with antineuraminidase serum.

    Original languageEnglish
    Pages (from-to)299-305
    Number of pages7
    Issue number2
    Publication statusPublished - 30 Nov 1988


    • cosmid vector
    • recombinant DNA
    • Streptococcus pneumoniae
    • virulence factor, gene bank


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